APOE3 Christchurch modulates ß-catenin/Wnt signaling in iPS cell-derived cerebral organoids from Alzheimer's cases.

A patient with the PSEN1 E280A mutation and homozygous for APOE3 Christchurch (APOE3Ch) displayed extreme resistance to Alzheimer's disease (AD) cognitive decline and tauopathy, despite having a high amyloid burden. To further investigate the differences in biological processes attributed to APOE3Ch, we generated induced pluripotent stem (iPS) cell-derived cerebral organoids from this resistant case and a non-protected control, using CRISPR/Cas9 gene editing to modulate APOE3Ch expression. In the APOE3Ch cerebral organoids, we observed a protective pattern from early tau phosphorylation. ScRNA sequencing revealed regulation of Cadherin and Wnt signaling pathways by APOE3Ch, with immunostaining indicating elevated ß-catenin protein levels. Further in vitro reporter assays unexpectedly demonstrated that ApoE3Ch functions as a Wnt3a signaling enhancer. This work uncovered a neomorphic molecular mechanism of protection of ApoE3 Christchurch, which may serve as the foundation for the future development of protected case-inspired therapeutics targeting AD and tauopathies.

Microplastic ingestion in key fish species of food webs in the Southwest Atlantic (Marine Protected Area Namuncurá / Burdwood Bank).

Microplastics (MPs) are currently one of the main problems of marine pollution, being found in all environmental matrices. Due to their size, they can be ingested by organisms directly (from the environment) or indirectly (with their prey). The objective of this study was to analyze the occurrence, abundance, concentration, and chemical nature of MPs present in the gastrointestinal tract of two fish species, Patagonotothen guntheri and Patagonotothen ramsayi, both of which are key in the food web of the Marine Protected Area Namuncurá/ Banco Burdwood (MPA N/BB). The analyzed species presented high values of MPs per individual (MPs/ind.) and occurrence compared to other studies. P. guntheri tended to have a lower number of MPs/ind. and occurrence than P. ramsayi (P. guntheri: 2.50 ± 1.93 MPs/ind., 82.50  %; P. ramsayi: 3.93 ± 2.91 MPs/ind., 90.60  %). While fibers were the predominant MPs in both species, P. ramsayi had a greater number of fragments and a greater variety of MPs chemical composition than P. guntheri. The prevailing chemical composition was cellulosic material (cellulose and cellulose mixed with polyamide and polyester). Synthetic fibers and fragments such as polyester (PET), alkyd resin, polyurethane, polyethylene, polyacrylic fiber and poly(ethylene-co-vinyl acetate-co-vinyl chloride) were also found. Although both species have a generalist diet, the differences found may be due to the fact that P. guntheri has benthopelagic feeding habits while P. ramsayi has demersal-benthic. Our study is the first report on the presence and characterization of MPs in organisms relevant to food webs in the Southwest Atlantic.

Riboflavin and Eosin Y Supported on Chromatographic Silica Gel as Heterogeneous Photocatalysts.

The application of photocatalysis for organic synthesis, both in the laboratory and on an industrial scale, will depend on the achieving of good yields and the ease with which it can be applied. Selective irradiation of the photocatalyst with LED light has made it possible to activate the reactions easily, without the need for UV or heat filters. However, a common problem is the need to separate the photocatalyst from the reaction products through extraction and chromatography isolation processes. These procedures make it difficult to recover and reuse the catalyst, which is not compatible with scale-up applications. Photocatalysts attached to heterogeneous supports resulted in an alternative, which facilitates their removal and reuse. In this study, we use chromatographic silica gel as a low-cost heterogeneous support to bind photosensitizers such as Riboflavin or Eosin Y. The modified silica gel was analyzed by FTIR-ATR and diffuse reflectance UV-visible spectroscopy, thermogravimetric analysis, and optical microscopy. These hybrid materials have a suitable size for easy separation by decantation and were found to be photoactive against two photooxidation reactions. These easy-to-handle materials open the door to effective applications for photoinduced organic synthesis methods at medium to large scale.

Microplastics distribution in river side bars: The combined effects of water level and wind intensity.

Rivers are the main pathway for microplastics (MP) transport toward the ocean. However, the understanding of the processes involved in the deposition and mobilization of MP in rivers, specifically in sediment side bars (SB), remains very limited. The objectives of this study were: (i) to examine the effect of hydrometric fluctuations and wind intensity on the distribution of microplastics (MP < 5 mm) in the SB of large river (the Paraná River), (ii) to determine the characteristics of MP to infer their origin and fate, and (iii) to discuss potential similarities or differences between MP suspended in the water column and MP found in sediment. The SB and water column were sampled during the autumn, winter, and spring of 2018, and the summer of 2019 at different river discharges and wind intensities. >90 % of the MP items found were fiber of polyethylene terephthalate (PET; FT-IR analysis), the most common MP color was blue, and most were in the 0.5-2 mm size range. The concentration/composition of MP varied according to the river discharge and wind intensity. During the falling limb of the hydrograph when discharge is decreasing and sediments are exposed for short periods (13-30 days), MP particles transported by the flow were deposited on temporarily exposed SB, accumulating there in high densities (309-373 items/kg). However, during the drought, when sediments remained exposed for a long time (259 days), MP were mobilized and transported by the wind. During this period (no influence of the flow), MP densities significantly decreased on SB (39-47 items/kg). In conclusion, both hydrological fluctuations and wind intensity played a significant role in MP distribution in SB.

TLR9 activation is required for cytotoxic response elicited by baculovirus capsid display.

Although the baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) infects lepidopteran invertebrates as natural hosts, represents an efficient vector for vaccine development. Baculovirus surface display induces strong humoral responses against viruses and parasites. A novel strategy based on capsid display carrying foreign antigens in the AcMNPV particle further improved the immune response by eliciting CD8+ T cell activation. In this study, we analyze the intracellular mechanisms and signalling pathways involved in CD8+ T cell activation by capsid display. Our results show that baculovirus can attach to the cell surface, enter dendritic cells (DCs), transit within endocytic vesicles and escape to the cytosol for further degradation by the proteasome. We found that the availability of viral proteins, endosomal acidification, and proteasome activity are needed for efficient Major Histocompatibility Complex class-I presentation by baculovirus carrying Ovalbumin in the viral capsid. Importantly, we demonstrated with this strategy that the induction of cytotoxic T cells and IL-12 production by DCs are TLR9-dependent and STING-independent. Finally, our study shows differential intracellular processing for capsid and surface baculovirus proteins in DCs and highlights the role of different danger receptors during cytotoxic T cell priming through the capsid display delivery system, which could lead to improved baculovirus-based vaccines development.

Gastro Update Europe 2020.

This narrative review summarizes a selection of clinically important novel gastrointestinal developments, presented and discussed at the virtual Gastro Update Europe. The selected topics, fully referenced, reflect what the distinguished faculty considered of vital importance to be communicated to the astute busy gastro-hep clinician who is eager to stay abreast of important novel developments. Whenever appropriate, a personal comment or addition was added to further raise the educational value of this review. Given its narrative character, statements and conclusions are largely expert opinion based. A full reference list of the lectures is included.

Long-term Follow-up of a Randomized Clinical Trial Comparing Endovascular Revascularization Plus Supervised Exercise With Supervised Exercise Only for Intermittent Claudication.

OBJECTIVE: The goal of this study was to assess the long-term effectiveness of combination therapy for intermittent claudication, compared with supervised exercise only. BACKGROUND: Supervised exercise therapy is recommended as first-line treatment for intermittent claudication by recent guidelines. Combining endovascular revascularization plus supervised exercise shows promising results; however, there is a lack of long-term follow-up. METHODS: The ERASE study is a multicenter randomized clinical trial, including patients between May 2010 and February 2013 with intermittent claudication. Interventions were combination of endovascular revascularization plus supervised exercise (n = 106) or supervised exercise only (n = 106). Primary endpoint was the difference in maximum walking distance at long-term follow-up. Secondary endpoints included differences in pain-free walking distance, ankle-brachial index, quality of life, progression to critical limb ischemia, and revascularization procedures during follow-up. This randomized trial report is based on a post hoc analysis of extended follow-up beyond that of the initial trial. Patients were followed up until 31 July 2017. Data were analyzed according to the intention-to-treat principle. RESULTS: Median long-term follow-up was 5.4 years (IQR 4.9-5.7). Treadmill test was completed for 128/212 (60%) patients. Whereas the difference in maximum walking distance significantly favored combination therapy at 1-year follow-up, the difference at 5-year follow-up was no longer significant (53 m; 99% CI-225 to 331; P = 0.62). No difference in pain-free walking distance, ankle-brachial index, and quality of life was found during long-term follow-up. We found that supervised exercise was associated with an increased hazard of a revascularization procedure during follow-up (HR 2.50; 99% CI 1.27-4.90; P < 0.001). The total number of revascularization procedures (including randomized treatment) was lower in the exercise only group compared to that in the combination therapy group (65 vs 149). CONCLUSIONS: Long-term follow up after combination therapy versus supervised exercise only, demonstrated no significant difference in walking distance or quality of life between the treatment groups. Combination therapy resulted in a lower number of revascularization procedures during follow-up but a higher total number of revascularizations including the randomized treatment. TRIAL REGISTRATION: Netherlands Trial Registry Identifier: NTR2249.

Plastic Impacts in Argentina: a Critical Research Review Contributing to the Global Knowledge.

PURPOSE OF REVIEW: Plastic pollution research has experienced exponential growth in the last decade; however, Argentina concentrates more than 70% of their research in the last 4 years. This review compiles regional research on plastic pollution in water, soil, sediment, air and organisms in Argentina. It discusses current sampling, quantification, and plastics identification methodologies while analyzing levels, gaps, and opportunities. RECENT FINDINGS: Research in plastic pollution was mainly focused on the biosphere component (52.9%), followed by the hydrosphere component (29.4%), and finally the lithosphere component (17.7%), with less than 20% addressing multiple components simultaneously. Sixty percent of this research was focused around microplastics, and less than 20% have considered multiple plastic debris sizes. Marine coastal species from Argentina had higher levels of microplastics than organisms from other South American studies, while microfibers were identified in 100% of the freshwater organisms studied. The lowest microplastic concentrations were found in lakes and in the Paraná and La Plata rivers, while the maximum concentrations were found in Pampa´s streams. There was a lack of standardization in methodology and unit expression in studies of sediment microplastics, which hinders comparison between reports. Argentine scientists have created the national alliance called SciEnce for Plastic Impacts Argentina (SEPIA). SEPIA is a network which aims to systemize plastic pollution research, coordinate methodologies, and enhance relationships with decision-makers, NGOs, and the general public. A time gap was found between the designation of principal international multilateral agreements and the implementation of national regulations for plastic waste treatment, with a tendency to include advanced concepts as Extended Producer Responsibility and Circular Economy.

Microplastics pollution in the intertidal limpet, Nacella magellanica, from Beagle Channel (Argentina).

Microplastics (MPs) have been identified in diverse marine invertebrates; however, there are few studies on limpets. This study aimed to evaluate and compare the presence of MPs in Nacella magellanica among sites of the Beagle Channel with different degree of anthropogenic influence. Intertidal limpets were sampled in three sites: Ushuaia Bay (UB) (inside Ushuaia city), a site with high anthropogenic influence owing to population growth, and fisheries, industrial and port activities; Ensenada Zaratiegui (EZ) and Playa Larga (PL) (west and east of Ushuaia city, respectively), both sites with a certain degree of environmental protection and low anthropogenic influence. MPs were isolated from the whole soft tissue of individuals by oxidative digestions with H2O2 (30%; 1:10 w/v). MPs occurrence was 100% in organisms from UB and EZ, whereas 90% in PL. Limpets from UB showed 10 ± 6.69 MPs/ind and 2.22 ± 0.78 MPs/g ww, while in PL and EZ there were 2.90 ± 2.02 MPs/ind and 0.96 ± 0.94 MPs/g ww; and 5.60 ± 2.59 MPs/ind and 2.41 ± 1.47 MPs/g ww, respectively. Fibres were the most abundant MPs type, followed by fragments and films. Fibres and films were identified as semi-synthetic cellulose and, although the polymer matrix of fragments was masked by copper phthalocyanine, one of them could be identified as polystyrene. As expected, the highest abundance of MPs was found in UB. However, a lower abundance of particles was detected in PL (downstream of UB) than in EZ (upstream of UB). This could be due to the influence of both the main current of the Beagle Channel and fluvial discharges, reducing the buoyancy of MPs. More studies are needed to understand the contribution of regional and local features to the accumulation of MPs in these environments and their interaction with the biota.

Transcriptome and metabolome analysis of crGART, a novel cell model of de novo purine synthesis deficiency: Alterations in CD36 expression and activity.

In humans, GART [phosphoribosylglycinamide formyltransferase (EC 2.1.2.2) / phosphoribosylglycinamide synthetase (EC 6.3.4.13) / phosphoribosylaminoimidazole synthetase (EC 6.3.3.1)] is a trifunctional protein which catalyzes the second, third, and fifth reactions of the ten step de novo purine synthesis (DNPS) pathway. The second step of DNPS is conversion of phosphoribosylamine (5-PRA) to glycineamide ribonucleotide (GAR). 5-PRA is extremely unstable under physiological conditions and is unlikely to accumulate in the absence of GART activity. Recently, a HeLa cell line null mutant for GART was constructed via CRISPR-Cas9 mutagenesis. This cell line, crGART, is an important cellular model of DNPS inactivation that does not accumulate DNPS pathway intermediates. In the current study, we characterized the crGART versus HeLa transcriptomes in purine-supplemented and purine-depleted growth conditions. We observed multiple transcriptome changes and discuss pathways and ontologies particularly relevant to Alzheimer disease and Down syndrome. We selected the Cluster of Differentiation (CD36) gene for initial analysis based on its elevated expression in crGART versus HeLa as well as its high basal expression, high log2 value, and minimal P-value.

Endovascular Revascularization Plus Supervised Exercise Versus Supervised Exercise Only for Intermittent Claudication: A Cost-Effectiveness Analysis.

[Figure: see text].

Reduced purine biosynthesis in humans after their divergence from Neandertals.

We analyze the metabolomes of humans, chimpanzees, and macaques in muscle, kidney and three different regions of the brain. Although several compounds in amino acid metabolism occur at either higher or lower concentrations in humans than in the other primates, metabolites downstream of adenylosuccinate lyase, which catalyzes two reactions in purine synthesis, occur at lower concentrations in humans. This enzyme carries an amino acid substitution that is present in all humans today but absent in Neandertals. By introducing the modern human substitution into the genomes of mice, as well as the ancestral, Neandertal-like substitution into the genomes of human cells, we show that this amino acid substitution contributes to much or all of the reduction of de novo synthesis of purines in humans.

First report of microplastics presence in the mussel Mytilus chilensis from Ushuaia Bay (Beagle Channel, Tierra del Fuego, Argentina).

Microplastics contamination is reported for the first time on Mytilus chilensis from Ushuaia Bay, one of the most remote areas of South America. Our results demonstrate the occurrence of microplastics in all samples, with an average of 8.6 ±â€¯3.53 items per individual. Fibers and fragments were the most abundant, with a percentage of 77.91 and 17.44%, respectively. The average length of the fibers was 742.3 ±â€¯702.1 µm, while the average cross-sectional area of fragments was 1944.80 ±â€¯960.94 µm2. The polymers identified were polyamides, semi-synthetic cellulosic materials, and PVC copolymers. The number of microplastics per individual was greater than those reported in even more populated regions. Probably, the modifications included in the technique led to optimize the extraction of MPs, reflecting the detection of a greater number of particles. M. chilensis could play a role in an extensive evaluation of MPs in the Ushuaia Bay, providing information on the interaction of MPs and biota.

The CRISPR-Cas9 crATIC HeLa transcriptome: Characterization of a novel cellular model of ATIC deficiency and ZMP accumulation.

In de novo purine biosynthesis (DNPS), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase (EC 2.1.2.3)/inosine monophosphate cyclohydrolase (EC 3.5.4.10) (ATIC) catalyzes the last two reactions of the pathway: conversion of 5-aminoimidazole-4-carboxamide ribonucleotide [aka Z-nucleotide monophosphate (ZMP)] to 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR) then to inosine monophosphate (IMP). Mutations in ATIC cause an untreatable and devastating inborn error of metabolism in humans. ZMP is an adenosine monophosphate (AMP) mimetic and a known activator of AMP-activated protein kinase (AMPK). Recently, a HeLa cell line null mutant for ATIC was constructed via CRISPR-Cas9 mutagenesis. This mutant, crATIC, accumulates ZMP during purine starvation. Given that the mutant can accumulate ZMP in the absence of treatment with exogenous compounds, crATIC is likely an important cellular model of DNPS inactivation and ZMP accumulation. In the current study, we characterize the crATIC transcriptome versus the HeLa transcriptome in purine-supplemented and purine-depleted growth conditions. We report and discuss transcriptome changes with particular relevance to Alzheimer's disease and in genes relevant to lipid and fatty acid synthesis, neurodevelopment, embryogenesis, cell cycle maintenance and progression, extracellular matrix, immune function, TGFß and other cellular processes.

Atmospheric Photo-oxidation of Diethyl Carbonate: Kinetics, Products, and Reaction Mechanism.

The rate coefficient for the gas phase of diethyl carbonate with chlorine atoms has been determined at 298 K using a relative method, employing ethyl formate and ethyl acetate as reference compounds. The experimental value, (1.0 ± 0.2) × 10-11 cm3 molecule-1 s-1, is in good correlation with the one estimated by the SAR (Structure-Activity Relationship) method. The photo-oxidation mechanism of diethyl carbonate initiated by chlorine atoms was also studied at 298 K and atmospheric pressure as a function of the oxygen partial pressure. The main products identified by infrared spectroscopy were CH3CH2OC(O)OCHO, CH3CH2OC(O)OCH2CHO, CH3CH2OC(O)OC(O)CH3, CO2, CO, HCOOH, and CH3COOH. The results reveal that the oxidation process occurs by the abstraction of a hydrogen atom from the methyl (43%) and methylene (57%) groups. The relative importance of each reaction path from the primary radicals formed in photo-oxidation and the identity of CH3CH2OC(O)OCHO, CH3CH2OC(O)OC(O)CH3, and CH3CH2OC(O)OCH2CHO were determined using computational methods. The activation energy of reaction paths for the main oxygenated radicals formed during photo-oxidation was determined using Gaussian09 Program.

The CRISPR-Cas9 crADSL HeLa transcriptome: A first step in establishing a model for ADSL deficiency and SAICAR accumulation.

Adenylosuccinate lyase (ADSL) catalyzes two steps in de novo purine synthesis (DNPS). Mutations in ADSL can result in inborn errors of metabolism characterized by developmental delay and disorder phenotypes, with no effective treatment options. Recently, SAICAR, a metabolic substrate of ADSL, has been found to have alternative roles in the cell, complicating the role of ADSL. crADSL, a CRISPR KO of ADSL in HeLa cells, was constructed to investigate DNPS and ADSL in a human cell line. Here we employ this cell line in an RNA-seq analysis to initially investigate the effect of DNPS and ADSL deficiency on the transcriptome as a first step in establishing a cellular model of ADSL deficiency. We report transcriptome changes in genes relevant to development, vascular development, muscle, and cancer biology, which provide interesting avenues for future research.

Mandible strength and geometry in relation to bite force: a study in three caviomorph rodents.

The monophyletic group Caviomorpha constitutes the most diverse rodent clade in terms of locomotion, ecology and diet. Caviomorph species show considerable variation in cranio-mandibular morphology that has been linked to the differences in toughness of dietary items and other behaviors, such as chisel-tooth digging. This work assesses the structural strength of the mandible of three caviomorph species that show remarkable differences in ecology, behavior and bite force: Chinchilla lanigera (a surface-dwelling species), Octodon degus (a semi-fossorial species) and Ctenomys talarum (a subterranean species). Finite element (FE) models of the mandibles are used to predict the stresses they withstand during incisor biting; the results are related to in vivo bite forces and interspecific variations in the mandibular geometries. The study concludes that the mandible of C. talarum is better able to withstand strong incisor bites. Its powerful adducting musculature is consistent with the notorious lateral expansion of the angular process and the masseteric crest, and the enhanced cortical bone thickness. Although it has a relatively low bite force, the mandible of O. degus also shows a good performance for mid-to-strong incisor biting, in contrast to that of C. lanigera, which exhibits, from a mechanical point of view, the worst performance. The mandibles of C. talarum and O. degus appear to be better suited to withstand stronger reaction forces from incisor biting, which is consistent with their closer phylogenetic affinity and shared digging behaviors. The contrast between the low in vivo bite force of C. lanigera and the relatively high estimations that result from the models suggests that its adductor musculature could play significant roles in functions other than incisor biting.